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Conjugation of doxorubicin to cell penetrating peptides sensitizes human breast MDA-MB 231 cancer cells to endogenous TRAIL-induced apoptosis

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Abstract

Previous work from our laboratory has shown that coupling doxorubicin (Dox) to cell penetrating peptides (Dox–CPPs) is a good strategy to overcome Dox resistance in MDA-MB 231 breast cancer cells. We also reported that, in contrast to unconjugated Dox-induced cell death, the increase in apoptotic response does not involve the mitochondrial apoptotic pathway. In this study, we demonstrate that both Dox and Dox–CPPs can increase the density of the TRAIL receptors DR4 and DR5 at the plasma membrane and moderately sensitize MDA-MB 231 cells to exogeneously added recombinant TRAIL, as has already been shown for other chemotherapeutic drugs. Moreover, we show that Dox–CPPs, used alone, induce the clustering of TRAIL receptors into ceramide-enriched membrane lipid rafts, a property not shared by unconjugated Dox and that this process is due to the generation of ceramide during Dox–CPPs treatment. In addition, MDA-MB 231 cells were found to express TRAIL and we show that the increased apoptotic rate induced by Dox–CPPs is due to the sensitization of MDA-MB 231 cells to endogenous TRAIL. The capacity of Dox–CPPs to sensitize cancer cells to physiologic amounts of TRAIL suggests that, in addition to their efficiency in combination chemotherapy, these compounds might increase the response of tumor cells to cytotoxic lymphocyte-mediated killing via TRAIL.

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Abbreviations

ASM:

Acid sphingomyelinase

CPP:

Cell penetrating peptide

Dox:

Doxorubicine

FACS:

Fluorescence activated cell sorting

Fc-R1, Fc-R2:

Recombinant proteins composed of IgG Fc fused to TRAIL-R or TRAIL-R2, respectively

FITC:

Fluoresceine isothiocyanate

MCa:

Maurocalcine

MCaAbu :

Maurocalcine analogue with cysteine residues replaced with l-α-aminobutyric acid

MCD:

Methyl-β-cyclodextrin

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide

NAC:

N-acetylcysteine

PBS:

Phosphate buffered saline

Pen:

Penetratin

ROS:

Reactive oxygen species

TRAIL:

TNF-related-apoptosis-inducing-ligand

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Acknowledgments

We thank Dr. Eric Gulbins (Department of Molecular Biology, University of Duisburg-Essen, Germany) for kindly providing anti-ceramide antibody and Dr. Olivier Micheau (INSERM, U866, France) for kindly providing soluble TRAIL-R1-Fc and TRAIL-R2-Fc. SA acknowledges the support of the Ministère de l’enseignement supérieur, de la recherche scientifique et de la technologie (Tunisia) for financial support and the University of Monastir.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Unité 05/UR/09-09, Mécanismes Moléculaires et Pathologies, Faculté de Médecine de Monastir, Monastir, 5019, Tunisie

    Sonia Aroui, Souhir Brahim & Abderraouf Kenani

  2. INSERM U836, Grenoble Institute of Neuroscience, Calcium Channels, Functions and Pathologies, Site santé de la Tronche, BP 170, 38042, Grenoble Cedex 9, France

    Sonia Aroui & Michel De Waard

  3. Université Joseph Fourier, BP 170, 38042, Grenoble Cedex 9, France

    Sonia Aroui & Michel De Waard

  4. INSERM U749, Faculté de Pharmacie, 5 rue J.B. Clément, 92290, Châtenay-Malabry, France

    Jocelyne Hamelin & Jacqueline Bréard

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  1. Sonia Aroui
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  2. Souhir Brahim
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  3. Jocelyne Hamelin
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  4. Michel De Waard
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  5. Jacqueline Bréard
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  6. Abderraouf Kenani
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Correspondence to Abderraouf Kenani.

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Jacqueline Bréard and Abderraouf Kenani are senior Authors and contributed equally to this work.

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Aroui, S., Brahim, S., Hamelin, J. et al. Conjugation of doxorubicin to cell penetrating peptides sensitizes human breast MDA-MB 231 cancer cells to endogenous TRAIL-induced apoptosis. Apoptosis 14, 1352–1365 (2009). https://doi.org/10.1007/s10495-009-0397-8

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